Boyko Alex, Kovalchuk Igor
Department of Biological Sciences, University of Lethbridge, Lethbridge, AB, Canada.
Methods Mol Biol. 2010;631:49-62. doi: 10.1007/978-1-60761-646-7_6.
Restriction fragment length polymorphism (RFLP) is a difference in DNA sequences of organisms belonging to the same species. RFLPs are typically detected as DNA fragments of different lengths after digestion with various restriction endonucleases. The comparison of RFLPs allows investigators to analyze the frequency of occurrence of mutations, such as point mutations, deletions, insertions, and gross chromosomal rearrangements, in the progeny of stressed plants. The assay involves restriction enzyme digestion of DNA followed by hybridization of digested DNA using a radioactively or enzymatically labeled probe. Since DNA can be digested with methylation sensitive enzymes, the assay can also be used to analyze a methylation pattern of a particular locus. Here, we describe RFLP analysis using methylation-insensitive and methylation-sensitive enzymes.
限制性片段长度多态性（RFLP）是同一物种生物体DNA序列中的差异。RFLP通常在使用各种限制性内切酶消化后被检测为不同长度的DNA片段。通过比较RFLP，研究人员可以分析应激植物后代中突变（如点突变、缺失、插入和染色体大片段重排）的发生频率。该检测方法包括对DNA进行限制性酶切，然后使用放射性或酶标记探针与消化后的DNA进行杂交。由于DNA可用甲基化敏感酶进行消化，该检测方法还可用于分析特定基因座的甲基化模式。在此，我们描述使用甲基化不敏感和甲基化敏感酶的RFLP分析。
